Tuberculosis is one of the infectious

disease that caused by

Mycobacterium tuberculosis.
Reported by Robert Koch in 1882.
Systematics of the Genus Mycobacteria

1874 Hansen --- Bacillus leprae

1882 Koch --- Bacterium tuberculosis
1896 Lehmann & Neuman genus
Mycobacterium, Mycobacteriaceae,
Actinomycetales, Actinomyces
M. leprae, M. tuberculosis
1974 Bergys Manual of Determinating
Bacteriology, 8th ed: 31 species
Groups of Mycobacteria
A. Slow growers :
1. Mycobacterium tuberculosis complex

2. Non tuberculous mycobacteria (NTM)

= Mycobacteria Others Than TB (MOTT)

B. Rapid growers

C. Non-cultivable or
Grow very poorly in culture media
1). Mycobacterium TB complex
M. tuberculosis = M.TB
M. bovis
M. bovis BCG
A caprine (goat) variant or M. bovis
M. africanum
M. microti
M. canetti
 2). Mycobacterium Other Than
Tuberculosis ( MOTT )
- before atypical mycobacteria atau NTM
(Non tuberculous mycobacteria )
- > 30 strain
Gr I Photochromogens
Gr II Scotochromogens
Gr III Nonchromogens
Gr IV Rapid growers
- contoh :
- M. kansasi
- M. avium intracellulare
- M. smegmatis
 Properties Mycobacterium TB (1)

Straight or slightly curved rods,

2~40.3~0.5 m
Singly
or in small clumps (in specimens),
serpentine cords (in broth)
Does not grow on ordinary culture media,
but only on enriched media egg-based,
e.g. LOWENSTEIN JENSEN MEDIUM (LJ),
KUDOH, OGAWA, and BACTEC.
-
Properties Mycobacterium TB (2)
Grows slowly, generation time of cells in
best condition of culture: 13-20 hours
Acid-fast, non-motile, non-sporing

Large amounts of lipid in cell wall: mycolic

acid colonies: a buff color, dry breadcrum-
like.
Optimal growth temperature: 35-37C
CULTURE OF M.TUBERCULOSIS (LJ)
 Mycobacterium tuberculosis (3)

Obligate aerobes, improves growth in CO2

Survive in milk & in other organic materials
Very sensitive to UV, also-heat-sensitive
Susceptibleto alcohol, formaldehyde,
glutaraldehyde; Resistant to drying
Niacin production (+)
 Acid Fast Staining

This is an acid fast stain of Mycobacterium tuberculosis

Note the red rods--hence the terminology for MTB in
histologic sections or smears: acid fast bacilli.
= Although ZN (+) (Acid Fast Bacilli = AFB) and
or Culture (+) (mycobacterium strain) the
results does not means that spesimen should
be Mycobacterium TB,
because AFB (+) or mycobacterium strain (+)
the possibility may be :
1) M.TB complex ( 7 strain)
2) MOTT (more than 30 strain)
= But for test PCR (+) by using primer TB
the result must be M.TB.
 Test Fluorescent

Mycobacteria can also be stained with auramine and viewed with

fluorescence microscopy, in which acid fast bacilli now appear as
glowing yellow rods. This method is easier to use to screen for
mycobacteria and is the method routinely used in sputum
specimens sent to the laboratory.
 Cell wall of M.TB
1. Cord factor ( 6,6 dimycolyl
trehalose )
2. Sulfolipid ( 2,3,6,6 tetra acyl
trehalose 2- sulfose)
3. Wax - D (not true wax )
4. Phosphatides
5. Firmly bound lipid. (the remaining
of mycolic acid after extraction by
organics solution)
 Diagnosis of Tuberculosis
Stains for Acid-Fast Bacilli
Biopsy Procedures
Conventional Culture Techniques
Radiometric Assays (BACTEC)
Mycobacteria Growth Indicator Tube
(MGIT) culture system
Polymerase Chain Reaction (PCR)
Nucleic Acid amplification TestsNAA
 Diagnosis of M.TB used in Microbiology
Depart, Fac.Med.Hasanuddin University.

1. Stains for Acid-Fast Bacilli

1.1. Kinyoun Gabbet stain
1.2. Ziehl Neelsen stain
1.3. Kinyoun Gabbet modification stain
2. Conventional Culture Techniques
e.g. Lowenstein Jensen media, Ogawa etc.
3. PCR : Polymerase Chain Reaction
4. Tes Elisa ( by using cord factor antigen).
THE IMPORTANT VIRUSES THAT INFECTED
HUMAN RESPIRATORY TRACT

1. ORTHOMYXOVIRUSES
Influenza virus A, B ,C
2. PARAMYXOVIRUSES
2.1. Measles virus sistim Tropmed
2.2. Mumps virus sistim Tropmed
2.3. Respiratory syncytial virussistim
Tropmed
2.4. Parainfluenza virus
3. OTHER VIRUSES
3.1. Corona virus sistim Tropmed
3.2. Rhino virus
3.3. Adenovirus
PROPERTIES OF ORTHOMYXOVIRUSES AND PARAMYXOVIRUSES

PROPERTY ORTHOMYXOVIRUSES PARAMYXOVIRUSES

VIRUSES INFLUENZA A,B,C VIRUSES ParaInfluenza 1-4,

Respiratory syncyitial virus,
Measles virus, Mumps virus.
GENOME Segmented (8 pieces) Non segmented ss RNA
ssRNA negative polarity negative polarity, a single piece.

Virion RNA Yes Yes

Polymerase
CAPSID Helical Helical
SYMMETRY
ENVELOPE Yes Yes
SIZE/diameter 110 nm 150 nm
SURFACE SPIKES Hemagglutinin and Neuromi Hemagglutinine and Neuromini-
nidase on different spikes dase on the same spike.
REPLICATION NUCLEUS CYTOPLASMA
a.Genetic a.Often a. Rare
reassorment
b.Rate of antigenic b. Higher b. Low
change
c.Giant cell- c. NO c. Yes
formation
Influenza Virus
 SKEMA VIRUS INFLUENZA A

HA
M1

NS2 M2

NA

LIPID BILAYER
NP

Transcriptase complex
(PB1,PB2,PA ) NS1 in infected cells
Trimer HA, Tetramer NA
 1. INFLUENZA VIRUS
The term Myxo refers to interact with
mucins (glycoproteins).
Three major hemagglutinin component
(H1,H2,H3) and two neurominidase
component (N1 and N2) in human.
Viral isolation in eggs or cell culture.
Mutability of Influenza A produces antigenic
changes.
Subtypes based of on H and antigen N.
 Hemagglutinin is the component on viral
membrane that attached to a specific receptor
on a susceptible cell acts in viral attachment
Neurominidase play role in envelope fusion and
viral release.
Both H and N molecules are immunogenic.
Nucleocapsid assembly takes place in the cell
nucleus.
Three major hemagglutinin component
(H1,H2,H3) and two neurominidase component
(N1 and N2) in human.
Viral isolation in eggs or cell culture.
 Antibody against the hemagglutinin
neutralize the infectivity of the virus
and prevent disease.
Antibody against group-spesific
antigen which is located internally
does not.
Antibody against neurominidase does
not neutralize infectivity but does
reduce disease.( perhaps reducing
spread).
Neuraminidase (NA)
Neuraminidase adalah enzim glikoprotein
antigenik yang berfungsi untuk memecah
asam sialat (N-acetylneuraminic acid)
Pemecahan asam sialat penting pada
saat budding virus karena apabila asam
sialat tidak dipecah, hemagglutinin akan
berikatan dengan reseptor tersebut dan
akan menghambat proses budding virus.
Selain asam sialat pada membran sel
inang, neuraminidase juga memecahkan
asam sialat yang terdapat pada mukosa.
 Mutability of Infl. A produces antigenic
changes.
Subtypes based of on H and antigen N.
Subtlype changes known as antigenic drift.
Antigenic drift every few years with type A.
Major antigenic shift due to reassorment
New subtype may also develop mutation.
Major antigenic shifts correlate with
epidemics
Minor antigenic drifts allow maintenance in
population.
 Hemagglutinin (HA)
Hemagglutinin is the antigenik gliko-
protein on the envelope of Influenza virus.
Hemagglutinin means to agglutinate of
the red blood cells.
Hemagglutinin of virus could be
attachment to the receptor of sel host
N-acetyl neurominic acid or sialic acid
 Hemagglutinin (HA)
In Avian influenza, HA tend to attach of
2,3 sialic acid reseptor while HA in
Human Influenza tend to attach of
2,6 sialic acid receptor.
2,3 sialic acid receptor on Avian
epithel respiratory with cilia.
2,6 sialic acid receptor on Human
epithel tracheobronchial without cilia.
 The difference of spesifisitas
reseptor HA depend on amino acid
position 226,
Glicin on Avian influenza virus
Leucin on Human influenza virus
The infection can be occur if protease sel
host broke HA HA1 and HA2.
HA2 function as fusion of the envelope
with membrane endosom during proses
uncoating.
 Influenza A show two types of
antigenic changes :
1. Antigenic shifts refers to major antigenic
changes that result from the reassorment
of the eight nucleic acid segments that
constitute the viral genome.
2. Antigenic drift refers to minor antigenic
changes resulting from point mutation in
the viral hemagglutinins and neurominida-
se glycoprotein.
Influenza virus (Orthomyxovirus)
RNA, envelope,helix, 90-120 nm
 Diagnosis :
Virus isolation detect virus
Direct immunofluorescence
Immunoenzymatic detection
Serodiagnosis
A/Hongkong/I/68
 A/Hongkong/I/68. . .H3. . . .N2

A : Strain Influenza Virus

Hongkong: Occured in Hongkong
I : First time isolate virus
68 : The Year of isolate
 2. PARAMYXOVIRUSES
PROPERTIES PARAMYXOVIRUSES
Composed of one piece of ss-RNA
Envelope helical nucleocapsid
Virioncontain RNA dependent RNA
polymerase, which transcribe the
negative polarity genome into m-RNA.
A fusion protein that causes cell fusion,
in some cases, hemolysis.
The envelope is covered with spikes,
H and N.
 PARAMYXOVIRUSES
Paramyxovirus family contains four
important human pathogens :
1. Measles virus
2. Mumps virus
3. Respiratory syncytial virus
4. Parainfluenza virus
ENVELOPE SPIKES OF PARAMYXOVIRYSES

VIRUS H N FUSION PROTEIN

Measles virus + _ +

Mumps virus + + +

Respiratory _ _ +
syncytial virus
Parainfluenza virus + + +
 CLINICAL FEATURE OF CERTAIN RNA
ENVELOPED VIRUSES

Rash Giant Type of Immunoglobulin

Virus cell
occurs Vaccine Commonly used
formed

Influenza No No killed No

Respiratory No Yes None No

syncytial
Measles Yes Yes Live No

Rubella Yes No Live No

Rabies No No killed No
 2.1. MEASLES VIRUS
PROPERTIES MEASLES VIRUS (1) :
ss-RNA
Envelope helical symmetry
Virion has two types of envelope spike,
haemagglutinine and fusion protein.
Single serotype.
Humans are natural host.
Transmitted by respiratory droplets by
coughing and sneezing.
 PROPERTIES MEASLES VIRUS (2):

Incubation period 10-20 days.

diameter 150 nm
Kopliks spot, a tiny, punctuate,whitish
rash with an erythematous base that
appears on the buccal mucosa just
above the lower molar, is patognomonic
for measles, 24-36 hours before rash
appears.
 PROPERTIES MEASLES VIRUS (3)
Virion RNA polymerase transcribes the
negative strand genome into m-RNA.
Lifelong immunity occurs in individual
who have had disease.
The exanthematic stage is characterize
by the typical morbilliform rash, which
begins on the face and thorax and
spread peripherally, approximately 5
days.
Measles virus ( Paramyxovirus)
envelope, helix, 150 nm
 2.2. MUMPS VIRUS

Properties Mumps virus.

# Family Paramyxovirus
# Envelope helical symmetry
# ss-RNA negative sense genome
# Hemagglutinin and Neurominidase on the
same spike.
# Only one antigenic type is known.
Mumps virus ( Paramyxovirus)
RNA,envelope, helix, 150 nm
MUMPS DISEASE
2.3. RESPIRATORY SYNCYTIAL VIRUS
PROPERTIES RESPIRATORY SYNCYTIAL VIRUS (1)
Classified as a pneumovirus with in the
Paramyxovirus family.
Pneumovirus causing syncytium formation in cell
culture.
No hemagglutinin and No Neurominidase.
ss-RNA, non segmented, negative sense
F-glycoprotein also mediate syncytium
formation
PROPERTIES RESPIRATORY SYNCYTIAL VIRUS (2)
Two glycoprotein, G-glycoprotein mediate
virus attachment to host cell receptor and
Fusion, F-glycoprotein induce fusion
of the viral envelope with the host cell
surface to facilitate the entry.
 Two antigenic subgroups A and B of RSV.
Epidemiologic study that group A
more severe
RSV is more important respiratory virus in
infants.
Nosocomial infection reduced by careful
handwashing.
 2.4. PARA INFLUENZA VIRUSES
PROPERTIES PARA INFLUENZA VIRUSES (1)
There are four type of parainfluenza
viruses,Parainfluenza 1,2,3, and 4
Paramyxovirus family.
Envelope viruses.
Ss-RNA, non segmented, negative sense
Posses H and N.
RNA synthesis occurs in the cytoplasm.
 PROPERTIES PARA INFLUENZA VIRUSES (2)
Antigenic of the four serotype relatively
stable.
Significant antigenic shift or drift does not
occurred.
Parainfluenza 1 is the major cause of acute
laryngotracheitis in infants and young
children, but less severe in upper respiratory
ilness(URI).
 PROPERTIES PARA INFLUENZA VIRUSES (3)
Para influenza 2 is slightly less significant
than parainfluenza 1 or 3.
Parainfluenza 3 is a major cause of severe
lower respiratory disease in infants and
young children.
Parainfluenza 4 is assopciated with mild
upper respiratory ilness
 3.1. CORONA VIRUS
PROPERTIES CORONA VIRUS (1)
Envelope helical symmetry
ss-RNA positive sense RNA genome
Outer surface has Club shaped
glycoprotein spikes give the virus a
crown-like appearance.
Incubation period 2-5 days
 PROPERTIES CORONA VIRUS (2)
The envelope has a lipid bilayer.
Two important strains 229E and OC 43
In the year of 2002 Severe Acute
Respiratory Syndrome (SARS).
 CORONA VIRUS :

Corona virus , RNA, helix, 80-160nm

Berseluibung, penyebab SARS
 3.2. RHINO VIRUS
PROPERTIES RHINO VIRUS (1)
Picornavirus group
ss-RNA, positive sense RNA genome.
Diameter 20-30 nm
Optimun temperature 33C for invitro replication
The receptor for most Rhino virus is
glycoprotein intercellular adhesion
molecule 1 (ICAM-1).
 PROPERTIES RHINO VIRUS (2)
Growing on Human diploid fibroblast
Incubation period 2-5 days
caused Common cold
Over 100 antigenically distinct
Rhinovirus have been identified.
 3.3. ADENOVIRUSES
PROPERTIES ADENOVIRUSES (1)
Almost 100 different serotype,
49 affect to human.
Naked Icosahedral symmetry.
ds-DNA
Diameter 70-90 nm, Capsomer 252
Replication and assembly occur in the
nucleus.
Virions are release by cell destruction.
 PROPERTIES ADENOVIRUSES (2)
Potensial for prolonged infection without
disease.
Transmission by spread person to person
by aerosol droplets and can infect the eye
by direct contact.
Portal entry by upper respiratory mucosa.
Type of infection : Lytic infection, persisten
infection,transforming infection.
Diagnosis of adenoviruses by virus
isolation or by culture.
Adeno virus, DNA, icosahedral
No envelope,70-90 nm